Sushi repeat-containing protein X-linked 2 (SRPX2), a chondroitin sulfate proteoglycan, functions as a critical regulator in several types of malignancy. However, its expression and biological functions in esophageal squamous cell carcinoma (ESCC) remain unclear. Thus, the objective of this paper was to investigate the expression pattern and biological functions of SRPX2 in ESCC. Our results demonstrated that SRPX2 is highly expressed in human ESCC tissues and cell lines. Knockdown of SRPX2 significantly suppressed the proliferation, migration and invasion of ESCC cells, as well as prevented the epithelial-to-mesenchymal transition (EMT) process in ESCC cells. Furthermore, knockdown of SRPX2 increased the sensitivity of ESCC cells towards cisplatin. Exploration of the underlying mechanisms of its action showed that knockdown of SRPX2 sharply down-regulated the expression levels of β-catenin, cyclin D1 and c-myc in ESCC cells. In conclusion, these findings indicated that knockdown of SRPX2 inhibits cell proliferation and metastasis, and promotes chemosensitivity in ESCC cells through the inactivation of Wnt/β-catenin signaling pathway. Thus, SRPX2 may be a promising target molecular for the treatment of ESCC.